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. 2021 Jan 26;11:2229. doi: 10.1038/s41598-021-82055-0

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Characterization of a SARS-CoV-2 replicon. (A) Kinetics of luminescence signal. The CHO-K1 cells were electroporated with 5 μg of replicon RNA. Intracellular luminescence signals were measured at the indicated time points. The mean and standard error of two independent experiments are shown in this figure. (B) Kinetics of the RNA copy. The CHO-K1 cells were electroporated with 10 μg of either the replicon RNA or the non-replicative N gene mRNA. RNA copy numbers were subsequently measured using qRT-PCR. The results were expressed as relative RNA copy number compared to that at 2 hpt. Multiple t-tests were performed for determining the statistical significance. A p-value < 0.05 was considered to be statistically significant. NS not significant. (C) The detection of N protein by IFA. The CHO-K1 cell was electroporated with 5 μg of replicon RNA. The cells were fixed with 4% paraformaldehyde, followed by permeabilization with 0.5% Triton-X. The expression of N protein was detected using anti-N mAb and goat-anti-mouse IgG conjugated with Alexa Fluor 488. Nucleus was stained by DAPI. (D) The detection of NSP8 protein by IFA. The expression of NSP8 protein was detected using anti-NSP8 mAb and goat-anti-mouse IgG conjugated with Alexa Fluor 488.